免疫磁分离及免疫比浊分析对福氏志贺菌的自动快速定量检测

目的 以SPA包被的磁珠和富含SPA的金黄色葡萄球菌分别与福氏志贺菌的多抗血清结合,制备福氏志贺菌特异的多抗磁珠和抗体致敏的金黄色葡萄球菌作为协同凝集试剂与免疫比浊试剂,利用免疫比浊分析探索其在福氏志贺菌自动快速定量检测中的初步应用.方法 福氏志贺菌F1a免疫日本大耳白兔制备多抗血清,磁珠包被SPA后再与多抗偶联,制成多抗免疫磁珠.以该磁珠捕获模拟标本的福氏志贺菌,接种MH平板37℃培养18～24 h,菌落计数后计算免疫磁珠的特异富集作用.富含SPA的金黄色葡萄球菌(No 1800株)经甲醛灭活后,与福氏志贺菌的多抗血清结合,制备特异抗体致敏的金黄色葡萄球菌,作为协同凝集试剂和自动定量检测的免疫比浊试剂,在日立7060全自动生化分析仪编制程序进行自动检测并进行初步的方法学评价.结果 福氏志贺菌Fla免疫日本大耳白兔获得了高质量的多抗血清,特异性好,凝集效价达1:320;制备多抗免疫磁珠对福氏志贺菌具有一定的富集作用,菌落计数显示磁珠处理的细菌捕获效率约为30%;抗体致敏的金黄色葡萄球菌,作为协同凝集试剂和自动定量检测的免疫比浊试剂,在玻片上能与待测福氏志贺菌产生明显的凝集现象,在日立7060全自动生化分析仪上定量检测灵敏、特异、线性良好.结论以福氏志贺菌的多抗血清分别致敏SPA包被的磁珠和富含SPA的金黄色葡萄球菌,制备特异的多抗磁珠对福氏志贺菌有一定的特异捕获与富集作用,抗体致敏的金黄色葡萄球菌作为协同凝集试剂与免疫比浊试剂对福氏志贺菌的自动、快速、定量检测具有较好的效果,为病原菌的快速诊断和自动定量分析提供了新的思路.     

Morphology and the sperm penetration assay

OBJECTIVE: To assess the ability of Kruger morphology to predict the outcome of the sperm penetration assay (SPA). DESIGN: A retrospective review using univariate and multivariate analysis, t tests, and logistic regression was performed to examine the correlation between Kruger morphology and the optimized zona-free hamster egg sperm penetration assay (O-HESPA). SETTING: University hospital. PATIENT(S): One hundred fifteen private-practice patients who underwent semen analysis, including Kruger morphology and O-HESPA, as part of an infertility evaluation between 1997-2000 were retrospectively reviewed. INTERVENTION(S): Retrospective analysis of the sperm penetration assay. MAIN OUTCOME MEASURE(S): Univariate and multivariate analysis, Student's t test, and logistic regression were performed to analyze Kruger morphology, count, and viability and their relationship to SCI result. RESULT(S): Univariate analysis demonstrated a statistically significant correlation between SCI and sperm count, viability, and Kruger morphology. Multivariate analysis demonstrated statistically significant correlations between SCI and count and viability. There was no correlation between Kruger morphology and SCI. Logistic regression was performed on the SCI results, using count, Kruger morphology, and viability. Sperm count was found to be the only variable that was statistically significant with respect to SCI results. CONCLUSION: This study demonstrated that Kruger morphology assessment cannot be used to predict the results of SCI or replace it in the management of the infertile couple.     

Standardized phase angle indicates nutritional status in hospitalized preoperative patients

There is currently no criterion standard to assess nutritional status, and different methods have been used in hospitalized patients. The aim of this study was to investigate the agreement and the association between bioelectrical impedance analysis derived by standardized phase angle (SPA) and other methods used for the nutritional assessment of body composition, metabolic status, and functional status in surgical patients. The hypothesis was that the SPA is effective for evaluating nutritional status in surgical patients; therefore, it could be used when the application of other assessment methods is not possible. The sample consisted of 125 patients (20-94 years of age) before elective gastrointestinal or hernia repair. The participants were from the Surgical Clinic 1 at the University Hospital of the Federal University of Santa Catarina, Florianópolis, SC, Brazil. Nutritional status was evaluated during the preoperative period based on the triceps skinfold thickness, mid-arm circumference, body mass index, percent weight loss, nutritional risk screening 2002 (NRS 2002), subjective global assessment (SGA), and SPA. The agreement between the SPA and the other methods was assessed with the k coefficient. The agreement between the SPA and the methods of nutritional assessment that were investigated for diagnosing malnutrition was moderate for NRS 2002 and SGA, weak for percent weight loss, and poor for triceps skinfold thickness, mid-arm circumference, and body mass index. There was a significant association between SPA and both NRS 2002 and SGA (P < .001). Our results suggest that SPA is able to indicate the risk of nutritional deficiency in the patients assessed. However, good agreement between SPA and the methods investigated was not observed.     

Scintillation proximity assay in lead discovery

Background: Scintillation proximity assay (SPA) is a homogeneous scintillant bead-based platform for the measurement of biological processes and plays an important role in the identification of active chemical entities in drug discovery. Objective: The design and development of solid-phase SPA approaches are examined and compared with alternative non-radiometric fluorescence-based technologies. Methods: This review provides background on the principle of SPA and its application to biomolecular interactions from a variety of biological sources. Conclusion: The SPA approach is well suited to the demands of commercial high volume automation and assay miniaturization for target-based high-throughput screening campaigns on synthetic and natural product libraries as well as for benchtop characterization and confirmation studies. In the near future, innovations in the way SPA and fluorescence-based screening strategies are multiplexed will improve our comprehensive understanding of cellular system biology and dramatically advance the lead discovery process for the treatment of complex target-related disorders.     

集对分析在卫生装备评估中的应用研究

介绍了一种处理模糊不确定问题的新方法——集对分析原理，对我军卫生装备评估进行了研究，建立了综合评估的数学模型，使定性评估定量化、精细化、明朗化，提高了评估工作的可操作性和科学合理性。     

Totally Transumbilical Laparoscopic Cholecystectomy

A recently convened Consortium at the Cleveland Clinic agreed on the term Laparo-Endoscopic Single-Site (LESS) surgery to describe minimally invasive techniques that use a single incision to accomplish laparoscopic procedures. These procedures are done by using either a single port through one fascial incision or multiple ports placed through separate fascial incisions. Because of cost containment issues and the lack of widespread availability of a single port, we currently use multiple reusable ports placed through three separate fascial incisions via a transumbilical incision. As opposed to standard laparoscopic cholecystectomy, a deflecting laparoscope and one articulating instrument are utilized to improve the safety and ease of this procedure. Presented in this video are the steps necessary to perform a LESS cholecystectomy via a transumbilical incision with commercially available instruments. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

胶体金免疫渗滤法快速检测幽门螺杆菌血清抗体的研究和应用

目的建立一种简便、快速、准确的检测胃幽门螺杆菌(H.pylori)抗体的新型血清学诊断方法。方法用胶体金标记技术与渗滤法相结合,建立了快速胶体金免疫渗滤法(DIGFA),与ELISA对比检测145份H.pylori感染血清,并与病理学检查对比检测108份H.pylori血清。做阻断试验和IgG标准品最小蛋白含量的检出观察,对其敏感性、特异性做了分析并调查了216例不同年龄组的人群感染率。结果本法与ELISA相比,两者符合率为95.1％;与病理组织染色检查比较,两者符合率为88.8％。DIGFA可检出IgG标准品的最小蛋白量为10μg     

Production and Characterization of Monoclonal Antibodies against Bovine Luteinizing Hormone

Five mouse hybridoma cell lines producing monoclonal antibody against bovine luteinizing hormone (LH) have been established and the respective antibodies characterized by radioimmunoassay, immunofluorescence and immunoelectrophoresis. All antibodies belong to the IgG class and bind to staphylococcus protein A. Intraspecies cross-reactivity studies revealed no reaction with bovine follicle stimulating hormone (FSH). However, all antibodies showed partial cross-reaction with bovine thyroid stimulating hormone (TSH) suggesting a close conformational similarity between bovine LH and TSH. Studies on interspecies cross-reactivity (rat and human) showed that three of these five antibodies strongly react with rat LH but not at all with either rat FSH or rat TSH thus representing monospecific reagents for investigations concerning LH in this species. One of these three antibodies also strongly binds to human LH and to the same extent to human chorionic gonadotropin (CG) but not to human FSH or TSH. It was concluded that at least three different epitopes on the bovine LH molecule are recognized and that they are located on the β-chain of the hormone.     

ABC-ELISA法诊断肠螨病的研究

目的　探讨生物素 -亲和素酶联免疫吸附试验 (ABC -ELISA)在肠螨病诊断中的应用价值。方法　采集 4 8例肠螨病患者血清 ,用ABC -ELISA法检测血清中螨特异性抗体IgG ,并与葡萄球菌A -蛋白酶联免疫吸附试验 (SPA -ELISA)进行比较。结果　用ABC -ELISA法和SPA -ELISA法检测 4 8例肠螨病患者血清中的螨特异性抗体IgG阳性率分别为89 5 8% (43/ 4 8)和 5 6 2 5 % (2 7/ 4 8) ,两者比较 ,差异具显著性 (χ2 =13 5 0 ,P <0 0 1)。结论　ABC -ELISA法是实验室诊断肠螨病的一种有效方法     

Feasibility study on single-port access (SPA) laparoscopic cholecystectomy using transanal endoscopic microsurgery (TEM) instruments

Abstract

Transanal endoscopic microsurgery (TEM) is a procedure for the treatment of rectal villous adenoma and early rectal tumours. It is actually the first single port access surgery (SPA) as well as a true Natural Orifice Transluminal Endoscopic Surgery (NOTES) in standard use for the last twenty years. The use of this technique and instrumentation in cholecystectomy is revisited. A feasibility study was done on porcine model. Peri-umbilical approach with the articulated TEM instruments was used for cholecystectomy. An in vivo study was started after the successful feasibility study. Cholecystectomy was performed on three consecutive porcine models using TEM instruments. Median operation time was 26.6 min. The post-op mean wound length was 3.7 cm. One gallbladder was perforated during the operation and the others were delivered intact. The instrument length was noted and the difficulty in instrument handling was documented. SPA cholecystectomy is feasible with Transanal Endoscopic Microsurgery (TEM) instruments. The special TEM technique of manipulation within a confined space is ideal for single port surgery. The TEM rectoscope can be used as a reusable single port and the instruments can be used as well but modification is suggested.